17-alpha-aminoalkynyl-3-chloro 19-norsteroidal 3,5-dienes

ABSTRACT

13 - ALKYL - 17A - AMINOALKYNYL - 3 - CHLOROGONA - 3,5DIEN-17-OLS AND 17-ALKANOATES (1) AND THEIR SALTS ARE USEFUL AS TRICHOMONACIDES, FUNGICIDES, AMEBICIDES AND BACTERICIDES. COMPOUNDS (I) ARE PREPARED BY TREATING THE CORRESPONDING 13 - ALKYL - 3 - CHLORO - 17A - ETHYNLGONA-3,5DIEN-17-OL OR 17-ALKANOTE (II) WITH FORMALDEHYDE AND AN APPROPRIATELY-SUBSTITUTED SECONDARY AMINE.

United States Patent 01.

3,553,213 Patented Jan. 5, 1971 ABSTRACT OF THE DISCLOSURE l3 alkyl 17maminoalkynyl 3 chlorogona 3,5- dien-17-ols and 17-alkanoates (1) andtheir salts are usein] as trichomonacides, fungicides, amebicides andbactericides. Compounds (I) are prepared by treating the corresponding13 alkyl 3 chloro 17a ethynylgona-3,5- dien-17-ol or 17-alkanoate (II)with formaldehyde and an appropriately-substituted secondary amine.

This invention is concerned with new 17a-aminoalkynyl- 3-chloro19-n0rsteroidal 3,5-dienes. The compounds of this invention arebiocidallyactive, especially against trichomonads, amebae, fungi andbacteria and, in particular, Staphylococcus aureus.

Description of the invention.'Ihis invention contemplates compounds ofFormula 1:

wherein R is alkyl of from 1 to 5 carbon atoms; R is hydrogen oralkanoyl of from 2 to 5 carbon atoms; and Y is wherein A and B,independently, are alkyl of from 1 to 5 carbon atoms,monophenyl-substituted alkyl of from 1 t carbon atoms or, takentogether, form a divalent radical selected from )2( 2)m )2- R beinghydrogen or methyl and m a whole number of from 2 to 6 -CH CH C (R CH CHR being hydrogen, hydroxy, methyl or carboxy,

-CH CH N R CH CH R being selected from alkyl of from 1 to 5 carbon atomsor monohydroxy-substituted alkyl of from 1 to 5 carbon atoms, or

and non-toxic, acid-addition salts thereof.

Preferred compounds comprise a subgeneric family of Formula I wherein Ris methyl or ethyl, R is hydrogen or acetyl and Y is Special mention ismade of a number of particularly valuable embodiments of this invention.These are:

(11 3 chloro 17 (3 diethylamino 1 propynyl-l3-ethylgona-3,5-dien-1713-01 and its hydrochloride, and particularly theenantiomorphs thereof and their hydrochlorides;

dl 3 chloro l3 ethyl 17m [3 (4 methyllpiperazinyl)-l-propynyl]-gona-3,5-dien-l7-ol and its di hydrochloride,and particularly the enantiomorphs thereof and their hydrochlorides;

3 chloro 17a (3 dimethylamino 1 propynyl)-l3- ethylgona-3,5-dien-17-oland its hydrochloride;

3 chloro 13 ethyl 17 [3 (4 methyl 1piperazinyl)-1-propyny1]-gona-3,5-dien-17B-ol, acetate and itsdihydrochloride;

3 chloro 13 ethyl 174x (3 morpholino 1 propynyl)gona-3,5-dien-l7fi-ol,17-acetate and its hydrochloride;

3 chloro l7 [3 (2,2,6,6 tetrarnethyl piper-idino)-l-propynyl]-13-ethylgona-3,5-dien-l7fi-o1 and its hydrochloride;

3 chloro 13 ethyl 17a [3 (pyrrolidino) 1 propynyl] gona-3,5-dien-17-o1and its hydrochloride.

3 chloro 13 ethyl 17cc [3 piperidino 1 propynyl]gona-3,5-dien-l7-ol andits hydrochloride;

3 chloro 17a [3 (4,4 dimethyl piperidino)lpropynyl]-13-ethylgona-3,5-dien-17-01 and its hydrochloride; and

1 [3 (3 chloro 13 ethyl 175 hydroXygona-3,5-dien-17-yl)-2-propynyl]-isonipecotic acid and its hydrochloride.

When used herein and in the appended claims, the term alkyl of from 1 to5 carbon atoms contemplates lower hydrocarbon radicals, straight chainand branched, including, for example, methyl, ethyl, propyl, butyl andpentyl radicals. The term alkanoyl of from 2 to 5 carbon atomscontemplates lower alkanoyl radicals such as acetyl, propionyl, butyryland pentanoyl. The term non-toxic, acidaddition salts contemplates saltsof the basic compounds of Formula I. These salts can be used to isolatethe compounds and, in addition, are just as useful biocidally as thefree bases and in many instances, because of improved solubilitycharacteristics, they lend themselves advan tageously to the formulationof a broader range of biocidal compositions. Illustrative of thesalt-forming acids contemplated are inorganic acids such ashydrochloric, sulfuric, nitric, phosphoric and the like; and organicacids, such as acetic, malic, citric, aconitic, pamoic, and the like.

The 17a-aminoalkynyl-3-chl0ro l9 norsteroidal 3,5- dienes of Formula Iare prepared by treating an appropriately-substituted3-chloro-17ot-ethynyl l9-norsteroidal 3,5-dien-l7-ol or alkanoate ofFormula II with formaldehyde and an appropriately substituted amine:

AT\ l I-I-N a B wherein A and B are as above defined, under Mannichcondensation conditions, according to the following wherein R, R A and Bare as above defined.

The process is carried out under conditions which are not particularlycritical. Those skilled in the art Will immediately recognize it to be atype of Mannich reaction. In general, the acetylenic steroid II issuspended in an inert solvent, such as dioxane, and is treated with amixture of aqueous formaldehyde and the appropriate amine, preferably inthe form of an addition salt with an organic acid, such as acetic acid,or an inorganic acid, such as hydrochloric acid. A very small amount ofa metal salt catalyst, preferably cuprous chloride, can be added toimprove the yield. The reaction is conducted at a temperature of fromabout 25 C. to about 100 C. and preferably at about 5575 C. for a periodof time long enough to effect the reaction; 2040 hours at 5575 C., forexample, provides good yields in most cases. The product is recovered inany convenient manner. One means is to pour the mixture into ice andwater, render the suspension basic (as with dilute aqueous sodiumhydroxide) and extract the basic product with ether. Evaporating theether leaves the product as a residue. Alternatively, addition of anacid to the ether solution causes precipitation of the product as anacid-addition salt.

Starting materials of Formula II for preparation of the compounds ofthis invention are available or can be prepared by techniques availableto those skilled in the art. One means comprises treating thecorresponding 13- alkyl-17ix-ethynyl gon-4-en-3-ones of H. Smith,Hughes, Douglas, Wendt, Buzby, Jr., Edgren, Fisher, Foell, Gadsby,Hartley, Herbst, Jansen, Ledig, McLoughlin, Mc- Menamin, Pattison,Phillips, Rees, Siddal, Siuda, -L. Smith, Tokolics and Watson, J. Chem.Soc., 1964, 4472-4492 with oxalyl chloride. In one manner of proceeding,a quantity of 1 g. of the 13-alkyl-l7a-ethynylgon-4-en-3- one isdissolved in 25 ml. of dry benzene and stirred with ml. of oxalylchloride and 0.1 g. of oxalic acid for one hour. The organic solvent isevaporated at reduced pressure and the residue taken up in ether, Washedwith NaHCO solution and Water. The solvent is dried and evaporated toleave the l3-alkyl-3-chloro-17a-ethynylgona 3,5 dien-17-ol (II) as aresidue. It may be purified, if desired, by recrystallization fromalcohol, methanol or mixtures of methanol and water.

The compounds of Formula I of this invention have been found to possessbiocidal properties, mainly in that they are inimical in very smallamounts of the order of 7.8-1000 ,ug/ml. in aqueous media to the growthof fungi, protozoa (trichomonads), bacteria and the like. The instantcompounds are thus useful in biocidal compositions in a variety ofimportant fields of use. For example, they can be formulated and used inand fungicidally-trichomonicidallyand 'bactericidally-active industrialcleaning compositions, and in soaps and detergents intended forveterinary use and in biocidally-active Wash solutions to decontaminatepremises, pastures, animal cages and the like, which have been infectedwith microorganisms, particularly of the type mentioned. They Will beapplied according to end use as powders, solutions, suspensions and thelike, containing the active substance generally in concentrations of0.1% to 0.7% by weight, or even as much as 1%, 1.5%, 1.8% 2% and up toabout 5%. In Washing solutions for pastures and barns, the activecompounds of this invention will be used generally in the range of fromabout 0.02% to .25% by weight.

Although, in common with most organic substances with relatively highmolecular Weights, compounds of Formula I have limited solubility inwater, those skilled in the art will have no great difficulty informulating them into a wide variety of biocidally-active compositions.In general, standard techniques can be employed and, where necessary,advantage is taken of the ability of compounds of Formula I to formsalts, such as acidaddition salts, which have enhanced solubility inWater. The active compounds per se can be made up in dilute aqueoussolution. They can, in addition, where required to be made up into moreconcentrated formulations with solvents such as N,N-climethylacetamideor ethylene glycol dimethyl ether and the like. They can also beformulated as suspensions or solutions in an aqueous vehicle containingan organic co-solvent, such as for example, N,N-dimethylacetamide oralcohol. Also, aqueous vehicles containing emulsifying agents, such assodium lauryl sulfate, and relatively high concentrations, e.g., up toabout 5% by Weight, of the compounds of Formula I can be formulated byconventional techniques.

Illustrative of the biocidal properties of the compounds of thisinvention are results of tests to determine antiprotozoal, especiallytrichomonicidal, activity.

A 12.5 mg. portion of test material (as based on active moiety) is addedto 2.5 ml. of 1% phosphate buffer, pH 6. Further two-fold dilutions aremade in the same buffer. A 1 ml. volume of each dilution is transferredto small sterile screw cap assay tubes containing 3.8 ml. of Diamondmedium (formula furnished by American Type Culture Collection,Rockville, Md.) and 0.1 ml. of calf serum. Each assay tube is inoculatedwith 0.1 ml. of a 4872 hour culture of T richomonas vaginalis strainATCC No. 13972. A control assay tube containing no test material issimilarly inoculated. The assay tubes are gently shaken and thenincubated for 48 hours at 30 C. Following incubation the tubes aregently shaken and with the aid of a Pasteur pipette a drop is depositedon a Spiers- Levy eosinophil counting chamber. The chamber is examinedby phase contrast micro-scopy. The number of organlsms present in 1 mm.is multiplied by 5000 in order to express the count per ml. Thedifference in the number of organisms present in the control tube and inthe tubes containing the test material represents the relatrve potencyof the test material and is expressed as the percentage kill at thespecific dose level. Metronidazole may be used as a suitable positivecontrol test material.

I The results of testing illustrative members of this invention forTrichomonicidal activity are given in the examples hereinafter.

Also illustrative of the biocidal properties of the in stant compoundsare results of tests to determine antifungal activity, especiallyagainst pathogenic fungi, such as Hisloplasma capsulatum.

A 50 mg. portion of test material is solubilized or suspended in ml. ofsterile distilled water. Further two fold dilutions are made in the samesolvent. A 0.2 ml. volume of each dilution is transferred to a sterilestainless steel capped 13 x 100 mm. culture assay tube containing 1.8ml. of Brain heart infusion fortified with sheep blood; the agarinfusion is in a molten stage at 4748 C. The assay tube contents arewell mixed and the agar is allowed to solidify as a slant. Each slant isinoculated with the yeast phase of H istoplasma capsulatum ATCC No.11407 which has been grown for at least 96 hours in the same medium atC.; the growth of the inoculum slant is suspended in 2 in. steriledistilled water and a drop of the suspension delivered by a Pasteurpipette is used to inoculate each assay tube slant. The assay tubes areincubated for 6 days at 35 C. The assay tube containing the least amountof test material which completely inhibits growth is reported as theminimal inhibitory value and is reported in terms of g/ml.

The results of testing illustrative members of this invention forantifungal activity are given in the examples hereinafter.

An additional illustration of the biocidal activity of the instantcompositions is demonstrated in the results of tests to determineanti-bacterial activity, especially against Staphylococcus aureus.

Antibacterial screening is carried out by an agar serial dilutiontechnique. Distilled water is used as a vehicle. The stock solution isprepared at 10,000 ,ug./Inl. of substance in the vehicle. Two-folddilutions are made in sterile water. One ml. quanitities of eachdilution are incorporated in 9 ml. Seed agar in sterile petri dishes.The hardened surface is inoculated with test organisms and incubated 18hours at 35 C. Spectrum: B. subtilis (2 strains), S. aureus (3),Mycobacterium sp. Pseudomonas aeruginosa, E. coli (2), S. partyphi,Brucella bronchiseptica, L. casei, S. faecalis. The endpoint is reportedas a minmal inhibitory concentration (MIC) expressed in g./ml.; theleast amount of test substance that will completely inhibit the testorganism. The results of testing illustrative members of this inventionfor antibacterial activity are given in the examples hereinafter.

Antiamebic screening is carried out according to a procedure similar toThompson, et ai., Antibio, and Chem. 6, 337- (1956). The test substanceis incorporated and diluted in the aqueous phase of a modifiedBoeck-Drbohlav diphasic medium fortified with rice starch. The medium isinoculated with polybacteria and a known number of trophozoites of E.histolytica NIH 200. After 48 hours of incubation at 35 C. thetrophozoites are counted. The results are expressed in MIC, definedhereinabove.

In the product of a total synthesis which has not included a suitableresolution stage the compounds prepared by the invention will be presentas racemates. Using a convention approved by Fieser and Fieser,Steroids, p. 336 (1959), the compounds designated as the di-forms arethe enantiomers corresponding in configuration to that of the naturalhormone estrone. The corresponding enantiomorphs are consequentlydesignated the l-forms and the racemates the dl-forms. Racemates will bedepicted by structural formulae which show only the enantiomorphs of thed-configuration. Often, an especially preferred embodiment will compriseeither the l-isomer or the d-isomer. For example,d-3-chloro-13-e'thyl-l7a- [3-(4-methyl-1-piperazinyl)-1-propyl] gona-3,5 -dien:17- 01 has 130 times the activity against Endameba histolyticaand 80 times the activity against T richomonas vaginalis shown by thel-enantiomorph; and the l-enantiomorph has 4 times the activity againstSaccharomyces cerevisiae shown by the dl-racemate and 16 times theactivity against T richophyton mentagrophytes shown by the d-form and130 times shown by the dl-racemate; and the l-enantiomorph has 4 timesthe activity against H istoplasma capsulatum shown by the dl-racemate.

Description of the preferred embodiments-The following examples show thepreparation of a number of compounds within the scope of this invention.They are merely illustrative and are not to be construed to limit thescope of the claims in any manner whatsoever.

EXAMPLE 1 dl-3-chloro-17a-(3-diethylamino-l-propynyl)-13-ethylgona-3,5-dien-17-ol and hydrochloride To a solution of 2.0 g. ofdl-3-chloro-13-ethyl-17aethynylgona-3,5-dien-17-ol, in 17.5 ml. ofdioxane is added 1.3 ml. of water, 0.8 ml. of 40% Formalin solution, 1.3ml. of acetic acid, 0.65 ml. of diethylamine, and 50 mg. of cuprouschloride. The mixture is stirred at 60 C. under nitrogen for 24 hours.The reaction mixture is then poured into ice-water and basified with 10%NaOH solution. The resulting precipitate is extracted with ether. Thecombined ether extracts are washed with water, saturated brine solutionand then dried (MgSO The clear ether filtrate containing the basic formof the product is acidified with isopropanolic-HCI. The resultingprecipitate is collected and recrystallized from ether/methanol toobtain 1.07 g. of material identified to be the title compound, M.P.2675-269" C. dec.

Analysis.-C H NOCl-HCl requires (percent): C, 69.01; H, 8.69; N, 3.09;Cl, 15.67. Found: (percent): C, 68.80; H, 8.78; N, 3.09; Cl. 15.88.

Tricho'monas vaginalis activity was 97% kill at 6.25 Lg/1111.; MICagainst Histoplasma capsulatum was Mgjllll. and antibacterial activityagainst B. subtilis was 7.81 ,ug/ml. and against S. aureus (4 strains)was 7.81, 15.6 and 15.6 ,ug./ml. resp.

EXAMPLE 2 1-3-chloro-17u-(3-diethylamino1-propynyl)-13-ethylgona-3,5-diene-l7-ol and hydrochloride To a solution of 4.0 g. of1-3-chloro-13-ethyl-17a-ethynylgona-3,5-dien-17-ol in 35 m1. of dioxaneis added 2.6 ml. of water, 1.6 ml. of 40% Formalin solution, 4.0 ml. ofacetic acid, 4.0 ml. of diethylamine, and 200 mg. of cuprous chloride.The mixture is stirred at 70 C. for 24 hours under nitrogen, then addedto ml. of ice water and basified with 10% sodium hydroxide solution. Theresulting precipitate is extracted with chloroform and the combinedextracts are filtered, washed with saturate brine solution and dried (NaSO The mixture is filtered through filter aid, and the chloroform isreduced to a small volume in vacuo. The residue is dissolved in ether,and the clear solution acidified with a 5 N isopropanolic HCl. Theresulting precipitate is collected and reprecipitated frommethanol-ether to obtain 3.9 g. of product, M.P., 252-260 C., dec., k242 I'D/l. (e=20,900),

(c:1% CHCl AnaIysis.-C H NOCl-HC1 requires (percent): C, 69.01; H, 8.69;N, 3.09; Cl, 15.67. Found (percent): C, 69.16; H, 8.64; N, 3.12; Cl,15.88.

'EXAMPLE 3 dl-3-chloro-13-ethyl-17u-[3-(4-methyl-1-piperazinyl)-1-propynyl]gona-3,5-dien-17-ol and dihydrochloride To a solution of 2.0 g.of dl-3-chloro-13-ethyl-17a-eth ynylgona-3,5-dien-17-ol in 17.5 ml. ofdioxane is added 1.3 ml. of water, 0.8 ml. of 40% Formalin solution, 2.6ml. of acetic acid, 0.7 g. of N-methylpiperazine, and 50 mg. of cuprouschloride. The mixture is stirred under nitrogen at 60 C. for 24 hours.The reaction mixture is then poured into ice-water and basified with 10%NaOH, and the resulting precipitate is extracted with ether. Thecombined ether extracts are washed with saturated brine solution anddried (MgSO The clear ether solution is acidified withisopropanolic-HCl. The product is collected and recrystallized frommethanol to obtain 0.4 of product, M.P. 260263 C. dec.,

AXE:

EXAMPLE 4 dl-3-chlorol7a- 3-dimethylamino-1-propynyl) -13-ethylgona-3,5-dien- 17-01 and hydrochloride To a solution of 2.0 g. ofdl-3-choloro-13-ethyl-l7aethynylgona-3,5-dien-17-01 in 17.5 ml. ofdioxane is added 1.3 ml. of water, 0.8 ml. of 40% Formalin solution, 1.3ml. of acetic acid, 0.3 g. of dimethylamine and 50 mg. CuCl. The mixtureis stirred at 60 C. under nitrogen for 24 hours. The reaction mixture isthen poured into icewater and basified with 10% NaOH solution. Theresulting precipitate is extracted with ether. The combined etherextracts are washed with water and saturated brine solution and thendried (MgSO The clear ether filtrate is acidified withisopropanolic-HCI. The product is collected and recrystallized frommethanol to obtain 1.1 g. of Product, M.P. 278-279" C. dec.,

Analysis.-C H NOCl-HCl requires (percent): C, 67.91; H, 8.31; N, 3.30;C1, 16.71. Found (percent): C, 68.06; H, 8.47; N, 3.10; Cl, 16.76.

Triclzomonas vaginalis activity was 98% kill at 25 ,ug./ml.; MIC againstHistoplasma capsulatum Was 62.5 ,ug./ml., against B. subtilis was 7.81,ug./ml., against S. aureus Smith was 62.5 ,ug./1'I1l. and 125 ,ugjml.against three other strains of S. aureus.

EXAMPLE 5 1-3-chloro-13-ethyl-17a-[3-(4-methyl-l-piperazinyl)-1-propynyl] gona-3 ,5 -dien- 17-01 and dihydrochloride A solution of 2.8g. of l-3-chloro-13-ethyl-17a-ethynylgona3,5-dien-17-ol, 25 ml. ofdioxane, 1.9 ml. of water, 1.15 ml. of Formalin (40%), 2.8 ml. ofglacial acetic acid, 2.8 ml. of N-methylpiperazine, and traces ofcuprous chloride is stirred for 24 hours at 70 C. (oil heat) undernitrogen. The reaction mixture is poured into water, made rbasic withdiluted sodium hydroxide and the free base extracted with chloroform andfiltered through filter aid. The chloroform layer is dried, and theprecipitate obtained on addition of isopropanolic hydrochloric acid tothe clear solution is dried over P at 80 C. to obtain 0.7 g. of thetitle compound; M.P. 252254.5 C., A 242 m 1 (20,600),

307, 3.45, 4.15, 6.20,, 3 ;+193 c. (c=0.9%, 95% EtOH).

Analysis.C H N OCl-H O requires (percent): C, 60.72; H, 8.12; N, 5.25;CI, 19.92; H O, 3.37. Found (percent): C, 60.53; H, 7.77; N, 5.46; Cl,20.34; H O; 6.44.

MIC against Saccharomyces cerevisiae was 7.8 ,tg./ml., againstTrichophyton mentagrophytes was 7.8 ,ug./ml., and against Histoplasmacapsulatum was 31.3 ,ug/ml.

EXAMPLE 6 dl-3-chlor0-13-ethyl-17a-[3 (4-methyl-l-piperazinyl)-1-propynyl]g0na-3,5-dienl7-0l, acetate and hydrochloride.

A suspension of 4.0 g. ofdl3-chloro-13-ethyl-17aethynylgona-3,5-dien-17-01, acetate, 35 ml. ofdioxane, 2.6 ml. of water, 1.6 m1. of Formaline (40% 4.0 ml. of

acetic acid, 4.0 ml. of N-methylpiperazine, and traces of cuprouschloride is stirred for 24 hours at 70 C. (oil bath) under nitrogen. Thereaction mixture is poured into water, made basic with diluted sodiumhydroxide and the free base extracted with ether. The precipitateobtained on addition of isopropanolic hydrochloric acid isrecrystallized from 20 ml. of methanol to obtain 1.8 g. of the product,M.P. 245-2475 c. x 241 111,, 24,950),

Analysis.C H N O Cl-2HCl-%H O requires (percent): C, 61.42; H, 7.82; N,4.95; Cl, 18.76; H O, 1.59. Found (percent); C, 61.20; H, 7.62; N, 5.39;Cl, 18.79; H O, 2.95.

EXAMPLE 7 dl-3-chloro-13 -ethyl-1 711- (3-morpholino-l-propynyl) gona-3,5-dien-17-ol, 17-acetate and hydrochloride To a solution of 1.0 g. ofdl-3-chlor0-13-ethyl-17a-ethynylgona-3,5dien-17-ol, acetate, in 9 ml. ofdioxane is added 0.7 ml. of water, 0.4 ml. of 40% Formalin solution, 1.0ml. of glacial acetic acid, 1.0 ml. of morpholine, and mg. of cuprouschloride. The solution is stirred and heated to 70 C. under nitrogen for24 hours. The reaction mixture is then poured into ice-water andbasified with 10% NaOH solution. The resulting precipitate is extractedwith ether, and the organic layer is Washed with saturated brinesolution and dried (MgSO The clear ether solution is acidified with 5 Nisopropanolic-HCI. On standing overnight at 10 C., needles form and theyare filtered off and dried over P 0 to obtain 0.86 g. of the' product,M.P. 203210 C. dec.; A 242 mg (24,500),

dl-3-ChlOIO-17OL-[3-(2,2,6,6tetramethylpiperidino)-1-propynyl]-13-ethylgona-3,5-dien-17-01 andhydrochloride A mixture of 1.1 g. ofdl-3-chloro-13-ethyl-17wethynylgona-3,5-dien-17,8-ol, 1.1 ml. of2,2,6,6-tetramethylpiperidine, 0.2 g. of paraformaldehyde, and 6 ml. ofdioxane was stirred and heated to C. under nitrogen for 24 hours. Thereaction mixture is then poured into ice water and basified with 10%sodium hydroxide. The resulting precipitate is extracted with ether, andthe ether solution is washed with brine, and dried (MgSO The clear etherfiltrate is acidified with isopropanolic-HCl, and the resultingprecipitate is filtered off to obtain 0.8 g. of material. Afterrecrystallizing from methanol-ether and leaching once with water, 0.4 g.of product was obtained M.P. 239-243 C., dec., x 242 mu (320,000).

Analysis.C H NOCl-HC1 requires (percent): C, 71.51; H, 9.10; N, 2.69;Cl, 13.62. Found (percent): C, 71.12; H, 9.03; N, 2.72; CI, 13.61. MICagainst Trichopllyton mentagrophytes was 7.8 ,tg./ml., and againstSacclzaromyces cerevisiae was 62.5 ,ug./ ml.

EXAMPLE 9 d-3-chloro-1 3-ethyl-1 7a- [3- (4-methyl-l-piperazinyl -1propynyl]-gona-3,5-dien-17-ol and dihydrochloride To a solution of 2.5g. of d-3-chloro-13-ethyl-17-ethynylgona-3,5-dien-17,8-ol in 23 ml. ofdioxane is added 1.6 ml. of water, 1.0 ml. of 40% Formalin solution, 1.6ml. of acetic acid, 0.9 ml. of N-methylpiperazine, and 60 mg. of cuprouschloride. The mixture is stirred and heated to 60-70 C. for 24 hrs. Thereaction mixture is then poured into ice water and basified with 10%sodium hydroxide solution. The resulting precipitate is extracted withchloroform. The combined extracts are then washed with saturated brinesolution and dried (Na SO The clear chloroform solution is evaporated to25 ml. and acidified with isopropanolic-HCl. The resulting precipitatewas collected and after reprecipitation from methanolether, the productis filtered off and dried over P in vacuo to obtain 2.2 g., M.P. 256260C., dec., k 242 m ((420,700), [a] -193 (c=1.0%; 95% EtOH).

Analysis. C H N OCl-2HCl-H O requires (percent): C, 60.72; H, 8.12; N,5.25; Cl, 19.92; H O, 3.37. Found (percent): C, 60.85; H, 7.77; N, 5.62;Cl, 19.72; H O, 3.58. MIC against Endameba histolytica was 7.8 g] ml.and against T richomonas vaginalis was 12.5 ,ug/ml.

EXAMPLE dl-3-chloro-13-ethyl-17a-[3-(pyrrolidino)-l-propynyl]gona-3,5-dien-17-ol and hydrochloride A solution of 3.0 g. ofdl-3-chloro-l3-ethyl-17a-ethynylgona-3,5-dien-17-ol, 1.95 ml. of water,1.2 ml. of 40% Formalin, 2.0 ml. of acetic acid, and 1.0 ml. ofpyrrolidine in 26.2 ml. of dioxane with 50 mg. of cuprous chloride isheated to about 70 C. and stirred under nitrogen for 24 hours. Themixture is poured into ice-water and basified with 10% sodium hydroxidesolution. The resulting precipitate is extracted 3 times with 200 ml. ofchloroform, and the combined extracts are washed with saturated brinesolution and dried over sodium sulfate. The clear chloroform solution isreduced to a small volume and acidified with isopropanolic-HCl thenadded to 300 ml. of ether. The resulting colored precipitate is removedby filtering and reprecipitating from methanolether. After threereprecipitations from methanol-ether, 0.9 g. of product in the form ofan off white powder is obtained, M.P. 275276 C. with decomposition,)tmax 242 my (621,600)

AKBI

max

EXAMPLE ll d-3-chloro- 17a- 3 -diethylamin0-l-propynyl)-13ethylgona-3,5-dien-17-ol and hydrochloride A solution of 4.0 g. ofd-3-chloro-13-ethyl-17a-ethynylgona-3,5-dien-17-ol, 2.6 ml. of water,1.6 ml. of 40% Formalin, 4.0 ml. of acetic acid, and 4.0 ml. ofdiethylamine in 35 ml. of dioxane with 200 mg. of cuprous chloride isstirred and heated to 70 C. for 24 hrs. The mixture is added toice-water and basified with 10% sodium hydroxide solution. The resultingprecipitate is extracted with chloroform. A heavy emulsion which formsis removed by filtering the mixture through filter aid and washing wellwith chloroform. The organic layer is washed with saturated brinesolution and dried over sodium sulfate. The clear chloroform solution isreduced to a small volume in vacuo and acidified with isopropanolic-HCI.The resulting precipitate is diluted with ether, and collected byfiltering. Recrystallization of the crude material from 150 ml. of 4:11methanol-ether mixture affords 2.3 g. of crystalline product, M.P.255257 C. with decomposition, )t 242 m (22,200)

[ ]D247 (c=l.0%, CHCl Analysis.C H NOCl-HCl requires (percent): C,69.01; H, 8.69; N, 3.09; Cl, 15.67. Found (percent): C,

KB Ami.

OHCl amines and the following 17a-aminoalkynyl-3-chloro-l3-ethylgona-3,5-dien-17-ols are obtained:

EXAMPLE 13 dl-3-chloro-1 3-ethyl-17-ethynylgona-3,5-dien-17fl-ol acetateA solution of 15.6 g. ofdl-3-chloro-13-ethyl-17-ethynylgona-3,5-dien-17B-ol in 250 ml. of aceticanhydride, ml. of acetyl chloride, and 12.5 ml. of pyridine is refluxedfor two hours. The organic solvent is removed under reduced pressure,and the crystalline residue is recrystallized from methanol to obtain15.9 g. of material, M.P. 138141 c. x 242 ma (3 22,300),

Analysis.C H O Cl requires (percent): C, 74.07;

H, 7.83; CI, 9.51. Found (percent): C, 73.69; H, 7.70; Cl, 9.78.

EXAMPLE 14 d-3-chloro-13-ethyl-17a-ethynylgona-3,5-dien-17-01 acetate Asolution of 5.0 g. of d-3-chloro-13-ethyl-17a-ethynylgona-3,5-dien-17-oiin a mixture of 80 ml. of acetic anhydride, 40 ml. of acetyl chloride,and 4 ml. of pyridine is heated to reflux for 2 hours. The organicsolvent is evaporated under reduced pressure (0.5 mm.). The residue iscrystallized from 50 ml. of methanol to obtain 3.2 g. of white needlesM.P. -157 C. with softening at 142.5 C., A 242 mu (e=23,600),

max.

Analysis.C H O Cl requires (percent): C, 74.07; H, 7.83; Cl, 9.51. Found(percent): C, 73.78; H, 7.49; Cl, 9.48.

EXAMPLE l5 dl-3-chloro-13-ethyl-17u-[3-piperidino-l-propynyl]gona-3,5-dien-17-ol and hydrochloride A solution of 3.0 g. ofdl-3-chloro-13-ethyl-17a-ethynylgona-3,5-dien-17-ol, 2.0 ml. of water,1.2 ml. of 40% Formalin, 2.0 ml. of acetic acid, 1.2 ml. of piperidinein 26 ml. of dioxane with 50 mg. of cuprous chloride is heated to 70 C.with stirring for 20 hours under nitrogen. Add the reaction mixture toice Water and basify with 10% sodium hydroxide solution. The resultingprecipitate is extracted with chloroform, and the combined organicextracts are washed with saturated brine solution and dried (Na SO Afterfiltering through filter aid, the clear chloroform solution isevaporated in vacuo to a small volume. Acidification with isopropanolicHC1 and dilution with ether affords an off-white precipitate. Thematerial is collected by filtration and reprecipitated frommethanol-ether to obtain 1.0 g. of product identified to be the titlecompound, M.P. 284-285 C. with decomposition, X 242 m (e=21,700).

Analysis.C H NOCl-HCl requires (percent): C, 69.81; H, 8.46; N, 3.02;C1, 15.26. Found (percent): C, 69.55; H, 8.61; N, 3.12; Cl, 15.04.

EXAMPLE 16 dl-3-chloro-17a-[3-(4,4-dimethylpiperidino)-1-propynyl]-13-ethylgona-3 ,5 -dien- 17-01 and hydrochloride A solution of 4.0 g. ofdl-3-chloro-13-ethyl-17a-ethyny1- gona-3,5-dien-l7-ol, 2.6 ml. of water,1.6 ml. of 40% Formalin, 4.0 ml. of acetic acid, and 4.0 ml. of4,4-dimethylpiperidine in 35 ml. of dioxane with 200 mg. of cuprouschloride is heated to 70 with stirring for 24 hours under nitrogen. Thereaction mixture is then added to ice water and basified with 10% sodiumhydroxide solution. The resulting precipitate is extracted with etherand the combined extracts are washed with saturated :brine solution anddried (MgSO The clear ether solution is acidified with 4 N isopropanolicHCl and the resulting precipitate is collected by filtration. Thematerial is recrystallized from MeOH-ether to obtain 3.1 g. of a productidentified to be the title compound, M.P. 285- 288 C. withdecomposition, k 241 m (e==2l,600).

Analysis.C H NOCl-HCl requires (percent): C, 70.71; H, 8.80; N, 2.84;Cl, 14.40. Found (percent): C, 71.08; H, 9.16; N, 2.87; C], 14.43.

EXAMPLE 17 dl- 1- [3 (3-chlorc 13 -ethy1-17,8-hydroxygona-3,5-dien-17-yl)-2-propynyl]-isonipecotic acid and hydrochloride A mixture of 4.0 g.of dl-3-chloro-13-ethyl-17a-ethynylgona-3,5-dien-17-ol, 2.6 ml. ofwater, 1.6 m1. of 40% Formalin solution, 4.0 ml. of acetic acid and 4.0g. of isonipecotic acid in 35 ml. of dioxane with 200 mg. of cuprouschloride is stirred and heated to 70 C. under nitrogen for 24 hours. Thereaction mixture is then added to ice water. The solid material iscollected by filtration, dried over P and suspended in methanol. Thesuspension is treated with 4 N isopropanolic HCl, and the resultingsolution is evaporated under reduced pressure to a small volume untilthe product begins to crystallize. The mixture is diluted with ether,and the material is collected by filtration. Two recrystallizations frommethanol-ether afford 0.6 g. of material identified to be the titlecompound, M.P. 254-257 C. with decomposition, A 241 m (e=21,200).

Analysis.C H NO Cl-HCl requires (percent): C, 66.13; H, 7.73; N, 2.75;CI, 13.94. Found (percent): C, 66.40; H, 7.95; N, 2.61; Cl, 13.78.

EXAMPLE 18 The procedure of Example 1 is repeated, substituting for thedl-3-chloro-13-ethylgona-17a-ethynyl-3,5-dien-l7- ol, appropriatelysubstituted 13-alkyl-3-chloro-17aethynylgona-3,5-dien-17-ols oralkanoates and the following 12 13-alkyl 3 chloro-17a-(3-diethylamino 1propynyl) gona-3,5-dien-17-ols and alkanoates are obtained:

onzom R: CH OH CH 2( 2 s a 3 z CH CH COCH COCH (CH CH EXAMPLE 19 Thesulfuric, nitric, phosphoric, acetic, malic, citric, aconitic and pamoicacid addition salts of 3-chloro-17a- (3-diethylamino 1propynyl)-13-ethylgona-3,5-dien- 17-ol (Example 1) are prepared bytreating the clear ether filtrate containing the basic form of theproduct with stoichiometric amounts of the respective acids in"isopropanol, then evaporating the solvents, leaving the addition salt asa residue.

,ug./ ml.

We claim: 1. A compound selected from those of the formula OR1 B Iwherein R is alkyl of from 1 to 5 carbon atoms; R is hydrogen oralkanoyl of from 2 to 5 carbon atoms; and Y is -CECCH3N wherein A and B,independently, are alkyl of from 1 to 5 carbon atoms,monophenyl-substituted alkyl of from atoms, or

-CH CH OCH CH and non-toxic, acid-addition salts thereof.

2. A compound as defined in claim 1 wherein R is methyl or ethyl, R ishydrogen or acetyl and Y is wherein 3. A compound as defined in claim 1which is 3-chloro- 17u-( 3 diethylamino 1 propynyl) 13 ethylgona-3,5-dien-17-ol. I

4. A compound as defined in claim 3 in the form of a hydrochloric acidaddition salt.

5. A compound as defined in claim 3 in the form of a l-enantiomorph,substantially free of the d-enantiomorph.

6. A compound as defined in claim 5 in the form of a hydrochloric acidaddition salt.

7. A compound as defined in claim 1 which is 3-chloro- 13-ethy1-17a-[3(4 methyl 1 piperazinyD-l-propynyl] gona-3,5-dien-l7-ol.

8. A compound as defined in claim 7 in the form of a dihydrochloric acidaddition salt.

9. A compound as defined in claim 1 which is 3-chloro- 170c-(3dimethylamino 1 propynyl)-13-ethylgona-3,5- dien-17-ol.

10. A compound as defined in claim 9 in the form of a hydrochloric acidaddition salt.

11. A compound as defined in claim 7 in the form of a l-enantiomorph,substantially free of the d-enantiomorph.

12. A compound as defined in claim 11 in the form of 5 a dihydrochloricacid addition salt.

13. A compound as defined in claim 1 which is3-chloro-13-ethyl-17a-[3-(4 methyl 1 piperazinyl)-1-propynyl]gona-3,5-dien-l7-ol, acetate.

14. A compound as defined in claim 13 in the form of a dihydrochloricacid addition salt.

15. A compound as defined in claim 1 which is3-chloro-l3-ethyl-17a-(3-morpholino 1 propynyl)gona- 3,5-dien-17ol,17-acetate.

16. A compound as defined in claim 15 in the form of a hydrochloric acidaddition salt.

17. A compound as defined in claim 1 which is 3 chloro 17oz[3-(2,2,6,6-tetramethylpiperidino)-1-propynyl]-13-ethylgona-3,5-dien-17-ol.

18. A compound as defined in claim 17 in the form of a hydrochloric acidaddition salt.

19. A compound as defined in claim 7 in the form of a d-enantiomorph,substantially free of the l-enantiomorph.

20. A compound as defined in claim 19 in the form of a dihydrochloricacid addition salt.

21. A compound as defined in claim 1 which is 3-chloro 13 ethyl 17a[3-(pyrrolidino)-1-propynyl] gona-3,5-dien-17-ol.

22. A compound as defined in claim 21 in the form of a hydrochloric acidaddition salt.

23. A compound as defined in claim 3 in the form of a d-enantiomorph,substantially free of a l-enantiomorph.

24. A compound as defined in claim 23 in the form of a hydrochloric acidaddition salt.

25. A compound as defined in claim 1 which is 3 chloro 13ethyl-17a-[3-piperidino-1-propynyl]gona- 3,5-dien-l7-ol.

26. A compound as defined in claim 25 in the form of a hydrochloric acidaddition salt.

27. A compound as defined in claim 1 which is3-chloro-17u-[3-(4,4-dimethylpiperidino) 1 propynyl]-13-ethylgona-3,5-dien-17-ol.

28. A compound as defined in claim 27 in the form of a hydrochloric acidaddition salt.

29. A compound as defined in claim 1 which is 1-[3-(3- chloro-13-ethyl17/3 hydroxygona-3,5-dien-17-yl)-2-p1'opynyl]-isonipecotic acid.

30. A compound as defined in claim 29 in the form of a hydrochloric acidaddition salt.

References Cited UNITED STATES PATENTS 3,264,330 8/1966 Moersch et a1.3,285,939 11/1966 Moersch et a1.

HENRY A. FRENCH, Primary Examiner US. Cl. X.R.

